Abstract:】Irritable bowel syndrome（IBS） is a common functional gastrointestinal disorder. Symptoms typically include abdominal dis－comfort or pain，as well as altered bowel habits. Stress is a certain adaptive self-protect response to stimulations of inside or outside. Long-term chronic stress can cause or aggravate IBS in many ways. The article will review effects of chronic stress on irritable bowel syndrome.

Abstract:Objective：To construct the recombinant adenovirus（Rad） with expressing inhibitor 1 of differentiation（Id1），and to estab－lish its animal model. Methods：AdEasy-Id1，the recombinant adenovirus plasmid which had been constructed previously，was pack－aged in HEK293 cells to obtain recombinant adenovirus Id1（RAd-Id1）. Then the titer of virus was determined by green fluorescent protein（GFP） labeling method. The over-expression efficiency of Id1 was measured by qRT-PCR and Western blot in HepG2 cells infected with RAd-Id1，and the effect of Id1 on proliferation of HepG2 cells was detected by MTS assay. Animal model of Id1 expressing in liver tissue was established through the way of tail vein injection of RAd-Id1. In order to determine the suitable load of RAd-Id1 virus for infecting the mice，viral load gradient exper－iment was divided into PBS control group，RAd-GFP control group and RAd-Id1 experimental group（n=5/group）. Time gra－dient experiment examined once every 5 days as a group，a total of 7 group（n=5/group），to explore the appropriate time. The expression of Id1 as well as the changes of alpha fetal pro－tein（AFP） and carcinoembryonic antigen（CEA） in liver tissues were determined by Western blot. The level of Id1 antibody and the concentrations of AFP and CEA in serum were detected by ELISA. Results：Recombinant adenovirus plasmid containing Id1 gene was successfully established，then the corresponding adenovirus were acquired and the measured titer was 1.5×1011 GFU/mL. Western blot and qRT-PCR analysis showed that Id1 mRNA and protein levels significantly increased in HepG2 infected with RAd-Id1 for 48 h and 72 h respectively（P<0.01）. MTS assay reported that the values in RAd-Id1 group were higher than those in NC group and RAd-GFP group after 96 h（2.00±0.06 vs. 1.18±0.05，2.00±0.06 vs. 1.10±0.07；F=51.350，P=0.000）；Western blot results showed：when compared with PBS group and RAd-GFP group，the RAd-Id1 group’s levels of Id1 as well as AFP and CEA in liver was higher. ELISA results showed：the concentrations of AFP and CEA in serum were positively correlated with the load of RAd-Id1（rs AFP=0.903，PAFP=0.014；rs CEA=0.964，PCEA=0.002）. There was no significant difference among these concentrations of Id1 antibody in serum of mice infected with different load of RAd-Id1（P>0.05）. Conclusion：The RAd-Id1 is successfully constructed，which serves as the vector tool to study the effect of Id1 on hepatic neoplasm at the cellular level. Mouse animal model of RAd-Id1 accompanied with the elevation of both AFP and CEA in liver also has been successfully established by tail vein injection，which suggests that the ectopic expression of Id1 may be related to the expression of AFP and CEA，but further studies will be required to determine how the mecha－nism works.

Abstract:Objective：To investigate the effects of exogenous SDF-1αdelivery or EPCs transplantation or SDF-1αcombined with EPCs transplantation on hepatic ischemia reperfusion injuryed rats. Methods：EPCs were extracted from bone marrow of LEWIS rats and cultured for days for phenotype identification with flow cytometry and functional identification with confocal laser. The orthotopic liver transplantation models were established in mice，which were divided randomly into five groups：A group，sham group；B group，surgery group；C group，SDF-1α group；D group，EPCs group；E group，SDF-1α+EPCs group. The rats were euthanased after 12 h reperfusion，and the liver tissues in each group were assayed with immunohistochemistry for liver regeneration and HGF ex－pression. The protein levels of VEGF，HGF，TGF-β，Bcl-2 and Caspase-3 in each group were assayed with Western blot，and the expression of VEGF，HGF and TGF-β in liver homogenates with ELISA. Results：The CD34 and CD133 positive rates of EPCs extracted from LEWIS rats were 78.32% and 70.76 respectively，and the binding capacity of ac-LDL and UEA-1 in EPCs accounted for over 80%. The number of EPCs（79.6±6.3） induced by SDF-1αwas significant higher than that in control group（36.2±2.9）（P=0.000）. In liver tissues of each group，the immunohistochemical scores in C and D groups of HGF secretion and liver regeneration were significantly higher than those in A and B groups（P=0.000），and the immunohistochemical scores in E group was significant higher than those in C and D groups（P=0.000）. The results of proteins ex－pression showed that the expressions of VEGF，HGF，TGF-β and Bcl-2 proteins were higher in C and D groups than in A and B groups（P=0.000）；the expressions of VEGF，HGF and TGF-β proteins were significantly higher in E group than in C and D groups（P=0.000），but there was no significant difference in Bcl-2 expression among C，D，E groups. The expressions of Caspase-3 protein were significantly higher in B，C，D groups than in A group（P=0.000），but the expression of Caspase-3 was significant lower in E group than in C and D groups（P=0.000）. The ELISA got the same results. Conclusion：Exogenous SDF-1α delivery combined with EPCs transplantation produces a superior protective effect on hepatic ischemia reperfusion injuryed rats，which could induce the re－generation of liver and reduce hepatocyte apoptosis.

Abstract:Objective：To investigate the immunogenicity and the immunotoxicity of interferon α-2a（IFNα-2a） and interferon α-2b（IFNα-2b） on Kunming mice. Methods：Kunming mice were randomly divided into six groups（eight mice in each group）：positive control group（treated with cyclophosphamide，P group），IFNα-2b high dose group（A group），IFNα-2b low dose group（B group），IFNα-2a high dose group（C group），low dose group（D group） and negative control group（treated with solvent，N group）. Mice were treated by intraperitoneal injection for 5 weeks. The serum concentration of IFNα，antibodies of interferon（Anti-IFN） and the im－munostaining result of alexin C3 were determined to evaluate the immunogenicity effect. In addition，the body weight，organ index，hematologic traits and histopathological examination were determined to evaluate the immunotoxicity effect. Results：For the concen－tration of serum IFN，group A（9.27±0.79） was higher than group C（7.80±1.26）（P=0.000）；group B（7.91±1.09） was higher than group D（5.97±0.98）（P=0.013）. For the concentration of Anti-IFN，group A（7.49±1.30） was lower than group C（10.31±0.69）（P=0.000）； the group B（6.18±1.02） was lower than group D（8.24±0.74）（P=0.013）. For the kidney immunohistochemistry，group A（0.51±0.00） was lower than group C（0.54±0.00）（P=0.000）；group B（0.42±0.00） was lower than group D（0.47±0.00）（P=0.013）. For the weight gain，group A was lower than group C（P=0.015）；group B was lower than group C（P=0.003）. There was no significant difference between the experimental group and the N group when comparing the kidney index，liver index，and spleen index. Levels of WBC，PLT，MCH，and LY in group A，B，C and D were higher than those in the N group，and were lower than those in the group P（P<0.05）. There was no significant difference among the IgA，IgM and IgG in the immunoglobulin test results. There was no significant pathological change in spleen slices in the experimental group. Conclusion：There is significant difference between IFNα-2a and IFNα-2b for immunogenicity. Moreover，there is no significant difference between IFNα-2a and IFNα-2b for immunotoxcity.

Abstract:Objective：To investigate effects of dexamethasone（DEX） on P311/transforming growth factor-β1（TGF-β1）/α-smooth muscle actin（α-SMA） signaling pathway in benign biliary stricture（BBS） model fibroblast（MF）. Methods：Two normal rabbits were treated with normal bile duct fibroblasts（NF），BBS model was established via discission and anastomosis of two rabbits and MF was acquired；then they were divided into five groups：NF，MF，MF plus different concentration of DEX（0.02，0.1 and 0.5 mg/mL） groups. After respectively incubated with 48 h，cell proliferation was investigated by CCK-8；Relative mRNA expression of P311，TGF-β1 and α-SMA were assessed by real-time PCR；Relative protein expression of TGF-β1 and α-SMA were investigated by western blot. Results：①optical density（A） in each group：0.331±0.002 in the NF group，0.533±0.005 in the MF group，0.487±0.003 in the MF+DEX 0.02 group，0.434±0.004 in the MF+DEX 0.1 group，0.381±0.004 in the MF+DEX 0.5 group；②P311 mRNA in each group：1.000 0±0.024 8 in the NF group，4.146 3±0.037 1 in the MF group，3.789 9±0.056 8 in the MF+DEX 0.02 group，3.566 1±0.148 1 in the MF+DEX 0.1 group，2.945 8±0.654 0 in the MF+DEX 0.5 group；③TGF-β1 mRNA in each group：1.000 0±0.034 6 in the NF group，2.647 9±0.048 5 in MF group，1.929 7±0.054 8 in the MF+DEX 0.02 group，1.678 2±0.080 2 in MF+DEX 0.1 group and 1.676 2±0.036 9 in the MF+DEX 0.5 group；④cells α-SMA mRNA in each group：1.000 0±0.056 0 in the NF group，4.025 7±0.065 9 in the MF group，3.644 9±0.196 4 in the MF+DEX 0.02 group，2.712 9±0.102 4 in the MF+DEX 0.1 group and 2.320 7±0.031 2 in the MF+DEX 0.5 group；⑤TGF-β1 protein values in each group：0.999 6±0.046 3 in the NF group，2.096 3±0.029 3 in the MF group，1.781 8±0.040 4 in the MF+DEX 0.02 group，1.531 4±0.032 5 in the MF+DEX 0.1 group，and 1.384 0±0.035 7 in the MF+DEX 0.5 group；⑥The number of α-SMA protein in each group：1.000 8±0.051 4 in the NF group，3.2314±0.1160 in the MF group，2.875 3±0.078 0 in the MF+DEX 0.02 group，2.262 8±0.059 8 in the MF+DEX 0.1 group and 1.940 8±0.093 7 in the MF+DEX 0.5 group. After 48 h of DEX intervention，cell proliferation was significantly inhibited and both the mRNA and protein expression of P311，TGF-β1 and α-SMA（all P<0.05，0.1~0.5 mg/mL） were significantly regulated down，with a dose-dependent manner. Conclusion：Mechanism of DEX on anti-BBS may be because it is relevant to modulation of P311/TGF-β1/α-SMA signaling pathway.

Abstract:Objective：To investigate the protective effects and mechanisms of total flavonoids（TF） on α-naphthylisothiocyanate-in－duced（ANIT-induced） cholestatic liver injury in mice. Methods：Sixty Kunming mice were evenly randomized into 6 groups，namely normal group，model group，biphenyl dimethylester group（0.15 g/kg），high-dose，middle-dose，and low-dose TF groups（1.2，0.6，0.3 g/kg）. Except for the blank control group，the acute liver injury model was established by intraperitoneal injection of ANIT，and went gastric lavage with total flavonoids extracted from Polygonum knotweed L. for 9 consecutive days. After fasting for 20 hours，the liver tissue and the blood from eyeballs were taken out，and the content of SOD，MDA and GSH in liver tissue，as well as the serum level of ALT，AST，LDH，TBA，TBIL were measured. Results：Compared with the model group,Serum ALT levels in high dose group mice of total flavonoids extracted from Polygonum knotweed L[（243.14±27.35） U/L]，Levels of AST[（345.02±24.01） U/L]，Levels of LDH[（505.97±68.56） U/L]，TBA[（112.02±17.16） nmol/mL]，TBIL[（15.08±4.09） nmol/mL] and GSH contents in the liver[（63.08±4.88） ?滋mol/mg] as well as MDA contents[（3.55±0.32） nmol/mg] were lower in high-dose TF group than in model group，The differ－ence was statistically significant（P<0.05）. Serum ALT levels in medium dose group mice[（330.56±24.74） U/L]，levels of AST [（438.60±32.51） U/L]，LDH[（597.90±66.90） U/L]，TBA[（144.62±16.37） nmol/mL]，TBIL[（27.36±6.51） nmol/mL] and GSH con－tents in the liver[（54.52±3.80） ?滋mol/mg] as well as MDA content[（5.65±0.35） nmol/mg] were also lower in middle-dose TF group than in model group,the difference was also statistically significant（P<0.05）. Conclusion：The total flavonoids extracted from P. knotweed exerts therapeutic effect on ANIT-induced liver in－jury with cholestasis in mice.

Abstract:Objective：To investigate the effect of B cell lymphoma 6 member B（BCL6B） on the proliferation and migration of human colorectal cancer cell line HCT116，and to explore its possible mechanism. Methods：Quantitative polymerase chain reaction（qPCR） and Western blot were used to detect and compare the endogenous expression of BCL6B in FHC and HCT116 cells. Liposome method was used to transfer the pcDNA3.1-BCL6B to HCT116 and the control group with empty vector was set. MTT assay，flow cytometry，wound healing assay and transwell chamber assay were used to detect the influence on proliferation，cycle and migration ability of cancer cell. Western blot was adopted to detect the expression of β-catenin，p-GSK3β，CyclinD1，MMP-7 and E-cadherin. Results：According to qPCR and Western blot，BCL6B expression was notably repressed in HCT116 cells as compared with FHC cells（P=0.017），and expression of BCL6B was significantly increased in HCT116 cells after transfection with pcDNA3.1-BCL6B for 48 h（P=0.000）. Compared with those in the control group，the OD492 value of HCT116 cells at the 4d time-point was decreased by 41.79% （P=0.040），and the percentage of cells in G1 phase was increased by 62.09%（P=0.001），48 h wound healing rate and transmembrane cells of HCT116 cells in the BCL6B group were significantly reduced（P=0.001）. The protein levels of β-catenin，p-GSK3β，CyclinD1 and MMP-7 of HCT116 cells in the BCL6B group were dropped by 65.66%（P=0.028），62.03%（P=0.001），60.87%（P=0.021） and 50.88%（P=0.030），respectively. E-cadherin was increased by 63.64%（P=0.018）. Conclusion：BCL6B inhibits the proliferation and migration of HCT116 cells，and its mechanism may involve in the inhibition of Wnt/β-catenin pathway.

Abstract:Objective：To search for genes related to the invasion and matastasis of hepatocellular carcinoma（HCC） with the gene trap－ping technology. Methods：Firstly，SMMC7721 cells were transfected with capture plasmid pU21 vector and then selected by G418 to set up cell lines. Secondly，transwell assay and wound healing assay were used to detect the invasion and migration abilities of these cell lines. Thirdly，5’-Full RACE，gene cloning experiment and gene sequencing experiments were used to identify what the genes were captured by pU21 vector in these cell lines. Results：About 300 cell lines stably transfected with pU21. Invasion and migration abilities of about 70 cell lines were obviously changed. One cell line，A554 was found that the gene captured by pU21 was long inter-genic non-protein coding RNA 52（LINC00052）. Conclusion：Gene trap technology is useful to find genes related to the invasion and matastasis of HCC.

Abstract:Objective：To study interaction sites of the glucose-regulated protein 78 kD（GRP78） with pre-S1 protein（PreS1） of hep－atitis B virus（HBV）. Methods：Being amplified by PCR，the open reading frame of GRP78 was subcloned into pW28 and expressed in Escherichia coli（E.coli） B834. The GRP78 protein was purified by nickel affinity chromatography column. Three truncated plasmids of HBV PreS1，including pGST-PreS1-X1，pGST-PreS1-X2，pGST-PreS1-X3，were expressed in B834 and then the bacterial protein was purified by GST affinity chromatography column. Interactions between GRP78 and PreS1 truncations were detected by pull down and microscale thermophoresis（MST）. Results：The recombinant plasmid of pW28-GRP78 was successfully constructed. The GRP78 protein and the fusion protein（GST-PreS1-X1，GST-PreS1-X2，GST-PreS1-X3） were obtained. The directly interaction between GRP78 and fusion protein were detected by pull down and MST，suggesting that the 1-65 amino acid residues of PreS1 play a key role for GRP78 binding to PreS1. Conclusion：Thanks to the molecular cloning and purified technology of protein purification，the fu－sion protein of GRP78 protein and the PreS1 truncated seg－ments is formed. And interaction sites of GRP78 protein and the PreS1 is primarily compared，providing preliminary data for future study.

Abstract:Objective：To investigate the clinical effect of plasma exchange （PE） plus continuous hemodiafiltration （CHDF） on pre-liver failure. Methods：Retrospective analysis was made on the clinical data in the 49 patients with pre-liver failure. Twenty-nine pa－tients were treated by PE combined with CHDF based on medical supporting treatment and 20 patients were treated by medical sup－porting treatment alone. Their therapenutic effects on biochemical parameters and clinical outcome were observed and compared be－tween the two groups. Results：Symptoms such as fatigue，poor appetite，and jaundice were relieved after non-biologic artificial liver support system（NBALSS）. The levels of C reactive protein（CRP），interleukin（IL）-6，alanine aminotransferase（ALT），total bilirubin （TBIL） after treatment decreased，and serum prothrombin activity（PTA） level increased in the PE plus CHDF group. While，all of the biochemical indictors after treatment differed significantly between the two group（all P<0.05）. Patients in the treatment group had a significantly higher improvement rate（ χ2=5.935，P=0.015） compared with that in the control group. Conclusion：The curative effect of PE plus CHDF based on medical supporting treatment is more remarkable than that of medical supporting treatment alone；therapeutic PE plus CHDF significantly increases the improvement rate of patients with pre-liver failure.

Abstract:Objective：To explore the clinical effect of ultrasound-guided transverses abdominis plane（TAP） block combined with in－travenous anesthesia in laparoscopic gastrointestinal surgery. Methods：Patients of 84 with laparoscopic gastrointestinal surgery were selected and divided into two groups according to the random number table method. The control group was treated with total intra－venous anesthesia，while the observation group was treated with ultrasound-guided TAP block on the basis of the control group. The visual analogue scale（VAS） score，postoperative recovery and adverse reaction were compared and analyzed. Results：VAS scores at 1，4，8，12 and 24 h in the observation group was significantly lower than those in the control group（P<0.05）. After the operation of 48 h，the first remedy pain time（11.67±1.23） h in the observation group was significantly longer than that in the control group （4.32±1.21） h，with statistically significant difference（t=27.607，P=0.025）. After 48 h of operation，the recovery time of the gastroin－testinal tract was without significant difference in both two groups（P>0.05）. As for pain in right shoulder，there were 12 patients （28.57%） in the observation group and 16 patients（38.10%） in the control group，without significant difference（ χ2=0.857，P=0.355）. None of the patients had adverse reactions such as respiratory depression，skin itching，vomiting，nausea and mental disorders. Conclusion：Implementing the full-time intravenous anesthesia combined with ultrasound-guided TAP block in the laparoscopic gas－trointestinal surgery can effectively reduce postoperative pain and prolong the first remedy analgesia time，with clinical pro－motion value.

Abstract:Objective：To diagnoze complex high anal fistula by combined use of MRI Ga fistula radiography and dynamic contrast enhanced MRI（DCE-MRI） and to explore its guiding significance for the diagnosis and treatment of high complex anal fistula. Methods：MRI confirmed 264 cases of high complex anal fistula complicated with small internal fistula，and uncleared branch fistula form December 10，2014 to December 10，2016. DCE-MRI check was firstly performed，than DCE-MRI and GA-DTPA 4 mL+ saline 16 mL in the fistula was soon performed through injection. Youden index and AUC test were performed to test the specificity and sensitivity of the results. Results：Fistulography+DCE-MRI clear displayed fistulography of high complex fistula，external fistula，branch fistula，and fistula T1WI；T2WI showed high signal. Abscess was irregular or horseshoe shaped，T1WI，slightly low signal，T2WI，high signal of fat suppression. The control results of surgery showed that fistulography+DCE-MRI obviously increased the diagnostic value for small fistula and small branch fistula. MRI scan for small fistula showed that Youden index was 31.60%，curve area was 0.576；MRI scan for small branch fistula showed that Youden index was -23.70%，and curve area was 0.176. DCE-MRI for small fistula showed that Youden index was 69.40% and curve area was 0.603；DCE-MRI for small branch fistula showed that Youden index was -4.80% and curve area was 0.201. Fistulography+DCE-MRI for small fistula showed that the Youden index was 87.80%，and curve area was 0.798；fistulography+DCE-MRI for small branch fistula showed that Youden index was 79.50% and curve area was 0.898. Conclusion：Fistulography+DCE-MRI have high clinical significance in the diagnosis and treatment of high complex anal fistula.

Abstract:Objective：To analyze the relationship between the liver pathological changes with the related serological indexes of chronic hepatitis B（CHB） complicated with pulmonary tuberculosis（PTB） in patients of mildly elevated alanine aminotransferase（ALT）（40 U/L

Abstract:Objective：To analyze the survival rate and the risk factors in patients with biliary atresia（BA） who underwent portoen－terostomy（Kasai） procedure. Methods：Totally 132 patients with biliary atresia who underwent Kasai procedure from Janunary 2007 to Janunary 2014 were recruited to this study. Data including age for Kasai procedure，presence of cirrhosis at the time of operation，use of steroids after operation and CMV infection were input in SPSS 19.0. Survival rates of the patients were calculated by life table. Kaplan-Meier method and COX proportional hazards model were used to analyze the suspected risk factors of survival. Results：During the 7-year follow up，35 survived and 97 were dead among all 132 subjects. The median follow up time of survival patients were 53.3（from 12 to 86.17） months. one-，two- and five-year survival rate were （32.0±6.0）%，（26.0±4.0）% and （24.0±4.0）% respec－tively. The survival rates for use of steroids after Kasai procedure or not were 43.9% vs 18.7%（P=0.027）. While the survival rates of cirrhosis at the time of the Kasai procedure or not were 18.9% vs. 36.2%（P=0.037）. Steroid and cirrhosis were regarded as main af－fecting factors for survival rate according to Kaplan-Meier method. However，only the usage of steroid was the solo independent risk factor for survival rates after Kasai procedure by COX proportional hazards model（RR=0.602，95%CI=0.377 to 0.964，P=0.034）. Conclusion：Kasai procedure is an effective treatment for pa－tients with BA，and proper usage of steroids after Kasai opera－tion seems to be one of the important treatments to elevate the survival rate.

Abstract:Objective：To explore the impact of CD57+ natural killer（NK） cells and macrophages CD68+ on the survival conditions of patients with primary esophageal cancer. Methods：Clinical data of 96cases of primary esophageal cancer from June 2013 to June 2015 in our hospital were retrospectively analyzed. Infiltration levels of CD57+ NK cells and macrophages CD68+ were evaluated by immunohistochemical staining method. The relationship between cells infiltrating level and the pathological characteristics was ana－lyzed. CD5+ NK and CD68+ macrophages relations with TNM staging，and the effect of cell infiltration on the survival conditions of primary esophageal cancer patients were analyzed. Results：The degree of infiltration of CD57+ NK cells in the epithelium of patients with primary esophageal carcinomawas 64.58% and that of CD68+ macrophages was 51.04%. There was no significant correlation be－tween the level of CD57+ NK cells and CD68+ macrophages in the epithelium of patients with primary esophageal carcinoma and the age，sex，tumor size，N stage，T stage and TNM staging（P>0.05）. There was no significant difference in CD57+ NK cells and CD68+ macrophage counts between CD57+ NK cells and CD68+ macrophages in primary esophageal cancer patients（P>0.05）. CD57+ NK cells and CD68+ macrophage counts were （8.35±5.36）/HP and （39.43±14.05）/HP respectively at stageⅠ-Ⅱ，significantly higher than those at stage Ⅲ-Ⅳ[（5.34±4.55）/HP，（20.18±9.62）/HP]（P<0.05）. The effect of cell infiltration on the survival of patients with primary esophageal cancer was analyzed by Cox model （Forward：LR，introducing α=0.05，excluding α=0.10）. The low level of death of CD57+ NK cells in tumor tissue of patients with primary esophageal cancer had a high risk of death（P=0.046，RR=0.794，95%CI=0.633 to 0.996）. The high level of death of CD68+ macrophages in tumor tissue of patients with primary esophageal cancer had a high risk of death（P=0.000，RR=1.925，95%CI=1.364 to 2.717）. Conclusion：CD57+ NK cells and CD68+ macrophages infiltration level can reflect the body’s immune status. CD57+ NK cells and CD68+ macrophage infiltration level has certain clinical value in determining the survival of patients with primary esophageal cancer.

Abstract:Objective：To effective evaluate the previous controlled clinical trials of bone marrow mesenchymal stem cells（BM-MSCs） transplantation for end-stage liver cirrhosis validity assessment. Methods：Wanfang database（1990.01-2016.05），CNKI（1990.01-2016.05），PubMed（1990.01-2016.05），Embase（1990.01-2016.05），The Cochrane Library（Issue 5，2016），Science direct（1990.01-2016.05），Medline（1990.01-2016.05） were retrieved for publication in all languages and relevant literature and experiments designed for randomized controlled trial（RCT）. All enrolled publication met the requirements according to the inclusion and exclusion criteria. Meta-analysis was performed by using Review Manager 5.3 software and end-stage liver disease（MELD） score，prothrombin time in－ternational normalized ratio（INR），aspertate aminotransferase（AST） were taken as the main analyzing indicators. Results：Eight refer－ences were enrolled including a total of 507 patients with liver cirrhosis（255 cases of control group，252 cases of BM-MSCs treatment group）. Compared with those of control group，BM-MSCs trans－plantation in patients with decompensated cirrhosis MELD score（MD=1.65，95%CI=2.8 to 0.50，P=0.005），AST（MD=0.26，95%CI=0.44 to 0.08，P=0.005），INR（MD=0.26，95%CI=0.44 to 0.08，P=0.005） significantly decreased after 1 month；BM-MSCs treatment effect were superior to that of control group after six months：MELD score（MD=1.65，95%CI=2.80 to 0.50，P=0.005），AST（MD=0.26，95%CI=0.44 to 0.08，P=0.005），INR（MD=0.26，95%CI=0.44 to 0.08，P=0.005）. Conclusion：Due to the potential of the immune regulation of BM-MSCs and its differentiation into liver cells，BM-MSCs can be used as a promising thera－peutic agent for liver cirrhosis. And the current study found that this therapy can be safe and effective in improving liver function. However，how to control different variables to optimize the treatment of cirrhosis of the liver is not clear，therefore，cirrhosis optimize treatment strategies need to be further explored in future clinical trials and research mechanisms.

Abstract:Objective：To report a case of isolated rectal tuberculosis（TB），and to made a literature review to improve the understanding of this rare condition. Methods：The data of clinical features，laboratory examinations，endoscopic manifestations，pathological charac－teristics，diagnosis and therapy of one patient with isolated rectal TB in our hospital were collected. Moreover，literature was summa－rized and compared. Results：The patient was hospitalized for abdominal pain，bloody purulent stool and emaciation. Endoscopy showed a huge isolated rectal ulcer. However，histopathologic biopsy found no caseating granuloma or acid-fast bacilli. Multiple acid-fast stool cultures and TB polymerase chain reaction（TB-PCR） assay were also negative. But the tuberculin purified protein derivative （TB-PPD） skin test was strongly positive. In addition，there was no evidence of parenteral tuberculosis infection. Moreover，after 3 months of diagnostic anti-TB treatment，the patient had a clinical remission. Endoscopy revealed the rectal lesion was much improved. The rectal ulcer was healed after a continuous anti-TB therapy of 15 months，no recurrence was occurred up to now. Conclusion：Iso－lated rectal TB is a rare disease without characteristic clinical，imaging and endoscopic features. The positive rates of caseating granu－loma or acid-fast bacilli found by histopathologic biopsy are rather low. However，TB-PPD test and diagnostic anti-TB treatmet are simple but effective strategies，which have important value for increasing the diagnosing rate of TB.

Abstract:

Abstract:Objective：To compare the expression，affinity purification and enzyme kinetics against representative inhibitors of full-length human phsphodiesterase 4B2 fused with 6His-SUMO on N-terminus（SF-hPDE4B2） and its 152-528aa truncate fused with 6His-SUMO on N-terminus（ST-hPDE4B2），to reveal the superiority of ST-hPDE4B2 over SF-hPDE4B2 as the target for high-throughput screen of hPDE4B2 inhibitors as potential anti-inflammatory agents. Methods：The two fusion proteins were expressed in Escherichia coli BL 21（DE3） and purified through Ni2+-NTA；phsphodiesterase activities were measured via alkaline phosphatase-coupled malachite green assay of phosphate in high-throughput mode. Results：After affinity purification，ST-hPDE4B2 exhibited spe－cific activity nearly 40-fold higher and activity yield over 100-fold higher than SF-hPDE4B2；SDS-PAGE and Western blot of 6His tag supported that there were a lot of polypeptides degraded in both of the purified fusion proteins. Inhibition constants of （R）-Rolipram and papaverine indicated that SF-hPDE4B2 mainly existed in the high-affinity state while ST-hPDE4B2 appeared princi－pally in the low-affinity state for （R）-Rolipram. Conclusion：ST-hPDE4B2 may have some advantages over SF-hPDE4B2 as the tar－get for the screening of inhibitors as potential anti-inflammatory agents.

Abstract:Objective：To establish the PCR method with capillary electrophoresis（PCR-CE） for the detection of rbcL gene of diatoms involved in drowning based on the specific primers for diatoms rbcL gene. Methods：With 29 microbes probably involved with drown－ing，a PCR-CE detection system with the primer for diatom rbcL gene labelled with fluorescent was established to verify the specificity and stability. The effectiveness of the PCR-CE method for the diatoms rbcL gene detection in corpses samples was evaluated by com－paring the results of MD-VF-Auto SEM. Results：Positive results（size=197 bp） were achieved with 6 diatoms species（Navicula sp.，Nitzschia sp.，Cyclotella sp.，Melosira varians，Synedra radians，Skeletonema） by the PCR-CE method，by contrast with negative out－comes for other species. The sensitivity of the above 6 species of diatoms DNA were 0.502，0.117，0.029，0.042，0.275，0.215 ng，respectively（20 μL PCR system）. 35 cases of corpses（3 corpses land deaths，2 waterborne corpse cases of non-drowning，30 drown－ing cases） were detected，the detection rate of diatoms in the lungs，liver and kidney of the corpses was 100%，63.3% and 53.3%，respectively. The total positive rate was 83.3%. When the diatom content >10/10 g，the total positive rate was 100% by MD-VF-Auto SEM method. The total positive rate was and 92.6% by PCR-CE，there was no statistical difference（ ?字2=2.077，P=0.491），otherwise，there was statistical difference（P<0.05）. Conclusion：Because of less sample consumption，the PCR-CE method based on the ND-rbcL primer has a good prospect in forensic application for diatom detection，and leads to easy popularization in drowning diagnosis.

Abstract:Objective：To select stable expression of reference gene and to explore the water channel protein 5 mRNA expression for drowning rat model in different time points. Methods：Drowning rat was modeled in different time points（0 min，n=6；15 min，n=6；4 h，n=6；16 h，n=6；48 h，n=6；72 h，n=6）. Total RNA was extracted from samples and reversed transcription was conducted for cDNA syn－thesis. Candidate reference genes were detected by real-time PCR. In order to screen out the most stable reference gene，geNorm and NormFinder was used to rank the expression of each gene. Then，AQP-5 mRNA expression was detected by qRT-PCR. Results：ACTB was the most stable reference gene. Our study showed that the expression of AQP-5 mRNA was downloaded except 72 h in this model（15 min/0 min=0.51，P<0.001；4 h/0 min=0.53，P<0.001；16 h/0 min=0.63，P<0.001；48 h/0 min=0.81，P<0.05；72 h/0 min=0.89，P>0.05）. Conclusion：AQP-5 mRNA expression has some positive meaning and improves forensic identification in drowning model.

Abstract:Objective：To investigate the preliminary sequence-activity relationship of Pseudomonas Aeruginosa arylsulfatase（PAAS） by site-directed mutagenesis and to recognize residues suitable for iterative-saturation-mutagenesis. Methods：Candidates of residues involved in catalysis were recognized by analysis of conformation of PAAS active site；mutants were generated by site-directed muta－genesis and characterized after recombinant expression and Ni2+-NTA affinity purification. Results：In comparison to the wild type，the mutagenesis of R55，M72，G138，D317 and K375 caused the significant decrease of catalytic efficacy；the mutagenesis of R55，D317 and K375 reduced substrate preference for 4-nitrophenylsulfate and/or thermostability，but the mutagenesis of M72 and G138 caused primarily the decrease of activity. Conclusion：M72 and G138 are suitable candidate sites for directed evolution by iterative-satura－tion-mutagenesis to improve mutant activity.

Abstract:Objective：To label the fulvestrant （an endocrine therapy drug for breast cancer） with the radioiodine. Methods：The fulves－trant was radioiodine labeled by improved chloramine T method. The paper chromatography was used to detect the labeling rate and the stability of the labeled compound. The column chromatography was used to detect the separation and purification of labeled com－pound. The changes in physicochemical property of the 131I-fulvestrant was detected by mass-spectrography；the combining ability and inhibiting ability of labeled compound to human breast cancer cells MCF-7（ER+） was detected by cell binding assay and MTT assay. Results：When the reaction lasted for 5 min at room temperature at the pH value of 7.5，the radiochemical yield of the radioio－dine labeling to fulvestrant was （62.34±1.80）%，radiochemical purity was （98.6±3.4）%，the half maximal inhibitory concentration（IC50） of 48 h was 35 μCi. The labeled compound was stable and its combining ability to human breast cancer cells MCF-7（ER+） was also retained. Conclusion：The radioiodine labeling to the fulvestrant is successful，and it may be used to develop a new drug for breast cancer.

Abstract:Objective：To analyze the expression profile variation of long non-coding RNA（lncRNA） in 3-O-C12-HSL-inhibted Mo-DCs maturation. Methods：Human CD14+ monocytes were separated by immunomagnetic beads and induced by hIL-4 and hGM-CSF. Experimental group was stimulated with 40 μmol/L 3-O-C12-HSL and 0.1 μg/mL LPS；control group was stimulated with 0.1% DMSO and model group was stimulated with 0.1 μg/mL LPS. lncRNA chip was used to analyze the differential expression among control group，model group and experimental group. Significant differe-ntial lncRNA（P<0.05） was analyzed and scatter plot was used to study the variance in 2 times differential lncRNA. Clustering analysis was used to study the 5 times differential lncRNA. Results：Comparing with those of control group and model group，1 386 lncRNA which have more than 2 times variation and significant differences by statistical analysis were selected out，including 548 up-regulated lncRNA and 838 down-regulated lncRNA. Meatime，153 lncRNA accounting for 11.04％ of all differential lncRNA had more than 5 times variation in differ－ence groups. Characteristic change were displayed among 2 times differential lncRNA in scatter plot analysis. The hierarchical clustering results displayed that 5 times lncRNA clustering efficient in different groups. According to GO analysis and mRNA pathway analysis，PPAR signaling pathway，calcium signaling pathway and NF-κB signaling pathway were enriched with differential lncRNA associated genes. Conclusion：lncRNA displaying characteristic change may involve in 3-O-C12-HSL-inhibted Mo-DCs maturation.

Abstract:Objective：In this study，CRISPR/Cas9 system was used to target cutting and restructuring of the mCherry red fluorescent reporter gene which inserted an inactivation sequence，and to evaluate the relationship between the length of homologous fragments and recombinant efficiency by the CRISPR/Cas9 system and mCherry reports genes. Methods：Cas9 expression plasmid pKD46-Cas was constructed by Golden Gate clone method and transfected into E.coli DH5α，and the arabinose induced Cas9 protein expression which was detected by Western blot assay. Using Golden Gate clone method，the mCherry red fluorescent reporter gene of pTac-mCherry-3guid plasmid was targeted insertion a random DNA sequence，whose flanking contain 0，40，80 and 120 bp homologues repeats respectivley with mCherry gene to construct the plasmids pQ0，pQ40，pQ80，pQ120. This gene insertion caused reporter gene inactivation. The pQ0，pQ40，pQ80 and pQ120 were transformed into E.coli DH5α which expressing Cas9，the corresponding bacteria solid medium were pQ0 group，pQ40 group，pQ80 group and pQ120 group. Then the ratio of the red colonies to the total colonies were statistic to analysis the recombination efficiency. Results：The result of PCR and sequencing analysis showed that pKD46-Cas and the vector pQ0，pQ40，pQ80，pQ120 were correctly constructed. Western blot test demonstrated that Cas9 prote in could successfully express in E.coli DH5α. There was no red colonies in pQ0 group. The difference of recombination efficiency of pQ0，pQ40，pQ80，pQ120 group was statistically significant（P<0.05），the length of homologous fragments was positive correlated to the recombination efficiency（r=0.792，P<0.01）. Conclusion：This study developed a new system based on CRISRP/Cas9 and mCherry fluorescent reports gene to detect the efficiency of homologous recombination. recombination may fail without the homologous fragment and the length of the homologous fragment from 0 to 120 bp had effect on the recombination efficiency. This system would be a new technology platform for further study of homologous homologous in prokaryotes.

Abstract:Objective：To clone uricase encoding gene in Candida utilis into the modified expression vector of pET-28a，and to make the uricase gene in Escherichia coli（E.coli） continuously express in secreted form with biological activity. Methods：To construct ex－pression vector pET-28a，RT-PCR method was used to delete the Lac O and Lac I sequence of pET-28a. The target gene was fused by the the N-terminal of uricase encoding gene and the signal peptide of protein A in Staphylococcus aureus. The fusion gene was cloned into the expression vector pET-28a，then transformed into E.coli DH5ɑ and expressed. The biological activity of expression product was identified and measured by SDS-PAGE gel electrophoresis. Results：The uricase gene in E.coli was continuously ex－pressed with biological activity in secreted form. Conclusion：As the normal flora in the gut，E.coli can be used to construct the ex－pression vector pET-28a-sUOX，which has secrete protein effect. Expression of uricase can be applied to the hyperuricemia model，which provide a basis for reducing uricase level through intestine.

Abstract:Objective：To evaluate the biomechanical properties of the self-designed absorbable lumbar interbody fusion cage in a sin－gle-level of cadaveric lumbar spine. Methods：Fifteen lumbosacral（L1-S1） specimens were randomly divided into three groups：auto－genous iliac bone group（n=5），shape memory polyurethane（SMPU） cage group（n=5），polyetheretherketone（PEEK） cage group（n=5）. Three different intervertebral implants were implanted by lateral lumbar interbody fusion after complete discectomy（L3-4） was per－formed. The axial failure load and ROM between preoperation and postoperation were tested by a mechanical testing machine. Results：The ROM showed that there was no observed statistical difference in the preoperative ROM of all directions among three groups（P>0.05）. In the autogenous iliac bone group，the ROM in all directions of postoperation was significantly higher than that of preoperation（P<0.05）. In the SMPU cage group，the ROM in all directions of postoperation was significantly lower than that of preop－eration（P<0.05）. In the PEEK cage group，the ROM in all directions of postoperation was significantly lower than that of preoperation（P<0.05）. The postoperative ROM of all directions was significantly lower in SMPU cage group and PEEK cage group than autoge－nous iliac bone group（P<0.05）. There was no observed statistical difference in the postoperative ROM of all directions between SMPU cage group and PEEK cage group（P＞0.05）. In axial compression testing，the failure load of PEEK cage group was significantly higher than that of SMPU cage group and autogenous iliac bone group（P=0.000），and the failure load of SMPU cage group was significantly higher than that of autogenous iliac bone group（P=0.000）. Conclusion：SMPU cage can meet initial stability and bear the lumbar physiological load for lumbar interbody fusion.

Abstract:Objective：To study its in vitro drug release of evodiamine nanocomplex oil-in-water nanoemulation（OECNE）and to inves－tigate its intestinal absorption. Methods：The dialysis was used to evaluate release profile，and the release behavior was studied by model and similarity factor f2. Single-pass intestinal perfusion was conducted to evaluate intestinal absorption. Results：The cumula－tive releases of OECNE were both about 3-fold that of EDA in 0.1 pH 1.2 hydrochloric acid solution and in pH 6.8 phosphate buffer solution，and OECNE was in accordance with Weibull model in two different solutions，and f2 also revealed OECNE have benefit in drug release. The intestinal absorption rate constant of OECNE was 2.32-fold，2.74-fold，3.36-fold，4.13-fold and 2.74-fold than that of free EDA in stomach，duodenum，jejunum，ileum and colon. The effective permeability of OECNE was 4.66-fold，3.70-fold，4.87-fold and 3.60-fold than that of EDA. Conclusion：OECNE was successfully used to enhance the absorption of EDA.

Abstract:Objective：To explore the drug release behaviors in vitro and pharmacokinetics in vivo of pyridostigmine bromide na－noemulsion（PBNE）. Methods：The high performance liquid chromatography was established to investigate the release behaviors in vitro and pharmacokinetic behaivors in vivo. Results：The release behaviors of PBNE in 4 media（pH1.2 HCl，pH6.8 PBS，pH7.4 PBS，pH7.8 PBS） was similar to those of pyridostigmine bromide（PB）. The MRT（0-24 h） of PBNE increased by 0.98 h than PB；the Cmax of PBNE was 1.55 times that of PB；the oral bioavailability of PBNE was 1.82 times that of PB. Conclusion：The release behaviors of PBNE in different release medium are similar in vitro. PBNE can change the pharmacokinetic behaviors in vivo；the oral bioavailabil－ity of PBNE is higher than PB and PBNE has a sustained-release effect in vivo.

Abstract:Objective：To develop a nucleic acid sequence-based amplification coupled ELISA method for detection of invasive as－pergillosis（IA）. Methods：In this experiment，the nucleic acid sequence-based amplification was combined to the high sensitive sys－tem of digoxingenin. The specific 18sRNA of Aspergillus was amplified by the isothermal digoxigenin（DIG）-labeling NASBA process，then the DIG-labeled NASBA amplicons were immobilized on streptavidin-coated microtiter plate by hybridized with a specific bi－otinylated DNA probe. The hybrids were colorimetrically detected by the addition of an anti-DIG antibodies linked to AIP and sub－strate（disodium 4-nitrophenyl phosphate）. The specificity，sensibility and the clinical diagnostic efficiency of this method were eval－uated by detecting the total RNA extracted from the ten-fold serially titrated Aspergillus spores，the RNA extracted from the compar－isons and 82 blood samples（32 positive cases，50 negative cases）. Results：The total RNA extracted from the ten-fold serially titrated Aspergillus spores was detected by NASBA-ELISA. The optical density value was linearly correlated with the logarithm of the spore amount（y=0.278x+0.119，R2=0.887），and this method could detect as little as 1 CFU aspergillums spore. The RNA extracted from the comparisons（E.coli，S.aureus，P.Aeruginosa，C.albicans，C.neoformans） and Aspergillus were detected by NASBA-ELISA. The results showed that NASBA-ELISA was highly specific for Aspergillus detection and did not cross-react with the non-target fungi or bacte－ria. The performance of NASBA-ELISA system was evaluated by detecting blood samples of 82 patients at high risk for IA. The sensitivity and specificity were 80.6% and 80.0%. The NASBA-ELISA A values were assessed by ROC analysis，area under the curve=0.755（95%CI=0.645 to 0.865）. Conclusion：The NASBA-ELISA method can detect Aspergillus sensitively and specifi－cally，so it provides a new way for the molecular epidemiologic investigation and early diagnosis of IA.

Abstract:Objective：to provide a scientific basis for the prevention and control of the hand-foot-and-mouth disease（HFMD） by di－viding different epidemic models according to the time series characteristics in 31 regions from 2010 to 2015. Methods：Time sequence features of HFMD incidence in different regions was extracted，including seasonal fluctuations，trends，autocorrelation and chaos，etc. Different time series epidemic patterns were divided by hierarchical clustering analysis. Results：Cluster analysis produced three types of epidemic patterns，namely，non-seasonal mode，single-season and multi-season seasonal pattern. Conclusion：For non-seasonal patterns，disease surveillance should be strengthened at any time to further analyzing the relevant factors. In the single-season and multi-season model，the appropriate preventive measures should be taken only in the high incidence period to reduce the possibility of a large area of epidemic disease.